fig2

Figure 2. Expression of paxillinS178A decreases cell spreading and directed cell migration. GFP-paxillin-wt and GFP-paxillinS178A MTLn3 cells were generated and three independent clones were used for further research. A: endogenous paxillin (red) colocalized with ectopically expressed GFP-paxillin-wt and GFP-paxillinS178A (green). Scale bar is 10 μm; B: cell clusters were detected using β-catenin (red) and GFP-paxillin (green) staining. Scale bar is 20 μm; C: cells were analyzed for cell adhesion (a). Cells were replated on collagen-coated plastic culture dishes. The number of attached cells was counted at different time points after replating. Columns show the mean of three independent experiments; bars show SE, ***P < 0.001. The spreading after 3 hrs of both wildtype and mutant cells was assessed using phase-contrast pictures (b); D: directed cell migration capacity was assessed using a woundhealing assay. The wound closure was measured at three different location in the wound after 24 h. The assay was repeated three times. Columns show the mean of three independent experiments; bars show SE, *** P < 0.001. All three adhesion related assays were demonstrating a defect in the GFP-paxillinS178A cells